The major area of study of this laboratory is the role of the epidermis as an immunological organ. We have found that epidermal Langerhans cells are derived from precursor cells in the bone marrow and play an essential role in many of the immunological reactions affecting the skin. We have demonstrated that when murine epidermal Langerhans cells are cultured for 2-3 days they become very potent antigen presenting cells for allogeneic and autologous T cells compared to freshly prepared Langerhans cells. We have therefore utilized cultured Langerhans cells for the generation of primary immune responses in resting unsensitized T cells. We have demonstrated that when cultured cells are modified with hapten, they can generate primary immune responses. The sensitized T cells thus generated respond preferentially to the same hapten in vitro. Another major focus of this laboratory has been the study of the function of class II MHC bearing keratinocytes which appear in humans and mice during cell-mediated reactions in the skin. Our studies demonstrate that these class II bearing keratinocytes induce specific immunological unresponsiveness in cloned T cells and in vivo in contact sensitization. We have also been studying the early cell and molecular events which occur after hapten painting of skin of nonsensitized mice to identify potentially important Langerhans cell and keratinocyte alterations. In response to hapten painting, activated Langerhans cells appear in vivo and there is a differential upregulation of epidermally-derived cytokine mRNAs. The cellular source of these cytokine mRNAs has been identified using cell depletion studies. We have found that IL-lbeta is the earliest cytokine to be activated. Furthermore, we have identified that IP-10, MIP-2 and IL-10 are produced by keratinocytes. We have also demonstrated that IL-1beta plays a critical role in the "activation" of Langerhans cells.